Abstract
We report on a differential scanning calorimetry study of native purple membranes under the following solvent conditions: 50 mM carbonate-bicarbonate, 100 mM NaCl, pH 9.5 and 190 mM phosphate, pH 7.5. The calorimetric transitions for bacteriorhodopsin denaturation are highly scanning-rate dependent, which indicates that the thermal denaturation is under kinetic control. This result is confirmed by a spectrophotometric study on the kinetics of the thermal denaturation of this protein. The calorimetric data at pH 9.5 conform to the two-state irreversible model. Comments are made regarding the information obtainable from differential scanning calorimetry studies on bacteriorhodopsin denaturation and the effect of irreversibility on the stability of membrane proteins.
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References
Bertazzon A, Tian GH, Lamblin A, Tsong TY (1990) Enthalpic and entropic contributions to actin stability: calorimetry, circular dichroism, and fluorescence study and effects of calcium. Biochemistry 29:291–298
Brandts JF, Hu CQ, Lin LN, Mas MT (1989) A simple model for proteins with interacting domains. Applications to scanning calorimetry data. Biochemistry 28:8588–8596
Brouillette CG, McMichens RB, Stern LJ, Khorana HG (1989) Structure and thermal stability of monomeric bacteriorhodopsin in mixed phospholipid/detergent micelles. Proteins 5:38–46
Brouillette CG, Muccio DD, Finney TK (1987) pH dependence of bacteriorhodopsin thermal unfolding. Biochemistry 26:7431–7438
Cladera J, Galisteoa ML, Duñac M, Mateo PL, Padros E (1988) Thermal denaturation of deionized and native purple membranes. Biochim Biophys Acta 943:148–156
Cladera J, Galisteo ML, Sabes M, Mateo PL, Padros E (1992) The role of retinal in the thermal stability of the purple membrane. Eur J Biochem 207:581–585
Conejero-Lara F, Mateo PL, Aviles FX, Sanchez-Ruiz JM (1991) Effect of Zn2+ on the thermal denaturation of carboxypeptidase B. Biochemistry 30:2067–2072
Edge V, Allewell NM, Sturtevant JM (1985) High-resolution differential scanning calorimetric analysis of the subunits of Escherichia coli aspartate transcarbamoylase. Biochemistry 24:5899–5906
Freire E, van Osdol WW, Mayorga OL, Sanchez-Ruiz JM (1990) Calorimetrically determined dynamics of complex unfolding transitions in proteins. Annu Rev Biophys Biophys Chem 19:159–188
Galisteo ML, Mateo PL, Sanchez-Ruiz JM (1991) Kinetic study on the irreversible thermal denaturation of phosphoglycerate kinase. Biochemistry 30:2061–2066
Goins B, Freire E (1988) Thermal stability and intersubunit interactions of cholera toxin in solution and in association with its cell-surface receptor ganglioside G m1 . Biochemistry 27:2046–2052
Hu CQ, Sturtevant JM (1987) Thermodynamic study of yeast phosphoglycerate kinase. Biochemistry 26:178–182
Jackson MB, Sturtevant JM (1978) Phase behavior of the purple membranes of Halobacterium halobium. Biochemistry 17:911–915
Kahn TW Sturtevant JM, Engelman DM (1992) Thermodynamic measurements of the contributions of helix-connecting loops and of retinal to the stability of bacteriorhodopsin. Biochemistry 31:8829–8839
Khorana HG (1988) Bacteriorhodopsin, a membrane protein that uses light to translocate protons. J Biol Chem 263:7439–7442
Klibanov AM, Ahern TJ (1987) Thermal stability of proteins. In: Oxender DL, Fox CF (eds) Protein engineering, Alan R. Liss, New York, pp 213–218
Kresheck GC, Lin CT, Williamson LN, Mason WR, Jang DJ, ElSayed MA (1990) The thermal stability of native, delipidated, deionized and regenerated bacteriorhodopsin. Photochem Photobiol 7:289–302
Lumry R, Eyring H (1954) Conformation changes of proteins. J Phys Chem 58:110–120
Lysko KA, Carlson R, Taverna R, Snow J, Brandts JF (1981) Protein involvement in structural transitions of erythrocyte ghosts. Use of thermal gel analysis to detect protein aggregation. Biochemistry 20:5570–5576
Maglova L, Guleva D, Chekulaeva L, Atanasov B (1990) A calorimetric study of white and purple membranes. Biochim Biophys Acta 1017:217–220
Manly SP, Matthews KS, Sturtevant JM (1985) Thermal denaturation of the core protein of lac repressor. Biochemistry 24:3842–3846
Morin PE, Diggs D, Freire E (1990) Thermal stability of membranereconstituted yeast cytochrome c oxidase. Biochemistry 29:781–788
Oesterhelt D, Hess B (1973) Reversible photolysis of the purple complex in the purple membrane of Halobacterium halobium. Eur J Biochem 37:316–326
Oesterhelt D, Stoeckenius W (1974) Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol 31:667–678
Privalov PL (1980) Scanning microcalorimeters for studying macromolecules. Pure Appl Chem 52:479–497
Privalov PL (1982) Stability of proteins. Proteins which do not present a single cooperative system. Adv Prot Chem 35:1–104
Privalov PL, Medved LV (1982) Domains in the fibrinogen molecule. J Mol Biol 159:665–683
Rigell WR, Freire E (1987) Differential detergent solubility investigation of thermally induced transitions in cytochrome c oxidase. Biochemistry 26:4366–4371
Ruiz-Sanz J, Ruiz-Cabello J, Mateo PL, Cortijo M (1992) The thermal transition in crude myelin proteolipid has lipid rather than protein origin. Eur Biophys J 21:71–76
Sanchez-Ruiz JM (1992) Theoretical analysis of Lumry-Eyring models in differential scanning calorimetry. Biophys J 61:921–935
Sanchez-Ruiz JM, Lopez-Lacomba JL, Cortijo M, Mateo PL (1988) Differential scanning calorimetry of the irreversible thermal denaturation of thermolysin. Biochemistry 27:1648–1652
Sanchez-Ruiz JM, Mateo PL (1987) Differential scanning calorimetry of membrane proteins. Cell Biol Rev 11:15–45
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Correspondence to: J. M. Sanchez-Ruiz
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Galisteo, M.L., Sanchez-Ruiz, J.M. Kinetic study into the irreversible thermal denaturation of bacteriorhodopsin. Eur Biophys J 22, 25–30 (1993). https://doi.org/10.1007/BF00205809
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DOI: https://doi.org/10.1007/BF00205809